细胞免疫和细胞凋亡在口腔扁平苔藓发病中的作用与意义

发布时间：2018-06-12 08:38

本文选题：口腔扁平苔藓 + 细胞免疫　；参考：《郑州大学》2014年博士论文

【摘要】：(1)口腔扁平苔藓(oral lichen planus,OLP)是临床上最为常见的慢性非感染性口腔黏膜病,是一种发生在皮肤及黏膜的慢性炎症性疾病。本研究通过对65例口腔扁平苔藓患者病变组织进行组织病理学检查证实其主要病理特征为:上皮过度角化或角化不全,棘层增生多见。上皮形状不规则,呈锯齿状。基底细胞层液化、变性,导致基底层角质细胞排列紊乱,基底膜界限不清,基底层角质细胞液化坏死。固有层中有密集的淋巴细胞浸润带,该浸润带主要分布在固有层的浅层,部分还可深入到固有层的深层或者粘膜下层。本研究65例口腔扁平苔藓患者中有23例在上皮的棘层、基底层或粘膜固有层可见圆形或卵圆形胶样小体,为均质性、嗜酸性,可能是属于凋亡的产物。5/65例出现上皮轻度不典型增生,表现为上皮细胞异乎于常态的增生,具体表现为增生的细胞大小不一,形态多样,核大而浓染,核浆比例增大,核分裂可增多但通常呈正常核分裂像。细胞排列较乱,细胞层次增多,极向消失。(2)应用免疫组化S-ABC法检测CD4+、CD8+T细胞在35例口腔扁平苔藓组织及10例正常口腔粘膜中的表达水平,结果表明:正常口腔粘膜上皮内几乎不表达CD4+、CD8+T细胞,仅基底层可见散在的CD4+、CD8+T细胞阳性表达。口腔扁平苔藓固有层显示中等量或大量淋巴细胞浸润,92.3%(60/65)的病例出现淋巴细胞浸润。口腔扁平苔藓组与对照组比较CD8+表达水平显著升高(P0.01),CD4+与对照组比较无显著性差异(P0.05),表明口腔扁平苔藓的发生可能与病变组织中细胞免疫改变有关。口腔扁平苔藓组CD8+T细胞主要位于固有层浅层,接近基底膜破坏处表达,CD4+T细胞主要于固有层深层表达。CD4/CD8比值下降,与对照组相比差异有统计学意义(P0.0 5)。(3)应用免疫组化S-ABC法检测Bcl-2和Bax在口腔扁平苔藓组及对照组中的表达水平。结果显示,Bcl-2主要表达于上皮基底层及棘层细胞的胞浆和胞膜,在正常口腔黏膜中,8/10例阳性(+)表达,阳性表达率80%;在口腔扁平苔藓的上皮组织中,17/35例阴性表达,3/35例在基底层和棘层呈零星散在表达,15/35例淋巴细胞浸润带处阳性表达增强,阳性表达率51.4%;Bcl-2在口腔扁平苔藓中阳性表达率与正常口腔黏膜比较无显著性变化(P0.05),但在淋巴细胞浸润带处表达增强。35例口腔扁平苔藓组织中,Bax阳性表达27例,阳性率77.1%,其中14例呈强阳性(+++)表达,9例为中度阳性(++),4例弱阳性(+),阳性细胞以基底层和棘层中下部最明显。10例正常人口腔黏膜中,3例弱阳性(+)表达,其余病例均为阴性(-),阳性表达率为30.0%。Bax在口腔扁平苔藓中阳性表达率和表达强度与正常口腔黏膜比较明显增高(P0.01)。(4)用TUNEL法(原位末端转移酶标记法)检测口腔扁平苔藓患者病变组织及对照组中细胞凋亡的情况,结果表明,65例口腔扁平苔藓病变组织口腔粘膜上皮层及固有层均可见凋亡细胞表达,核轮廓清楚、致密,呈棕褐色。口腔扁平苔藓病变组织的上皮细胞凋亡指数明显较正常对照组高(P0.01),凋亡细胞多位于基底层液化坏死处,表达率为85.7%(30/35)。而对照组中的凋亡细胞多位于表层、棘层及基底层,表达率为80%(8/10)。(5)双抗夹心酶联免疫吸附试验检测外周血血清TNF-a和IFN-γ含量,结果表明口腔扁平苔藓患者外周血血清IFN-γ和TNF-α含量均显著高于对照组(P0.05)。(6)采用单克隆抗体花环法对T细胞亚群进行识别与测定,结果发现,与健康组比较,20例口腔扁平苔藓患者CD8+显著升高(P0.05),CD4+与对照组比较无显著性差异(P0.05),CD4+/CD8+与对照组比较显著降低(P0.01),和免疫组织化学染色分析结果一致。(7)采用透射电镜技术观察发现口腔扁平苔藓患者口腔黏膜棘层胞质内充满张力原纤维,并变粗增多,排列紊乱,内织网扩张,内含不定形物,线粒体肿大及峭消失,游离核糖体增多。多数细胞器减少,细胞核团块状异染色质增多,核周间隙加宽,有的核旁空泡,细胞变形,桥粒结构破坏,细胞间隙明显扩大。基底细胞与基底膜半桥粒及细胞间桥粒破坏,部份细胞变形,排列紊乱,错位,水肿细胞间隙加大,空泡,变性,胞质内线粒体肿胀,峭明显减少,内质网扩张。细胞核增大,异染色质增多,凋亡细胞增多。凋亡细胞出现频率高于健康组。凋亡主要表现为染色质沿核膜边聚,且呈“C”形、新月形或团块状,并可见凋亡小体。结论1.口腔扁平苔藓的病理组织学特征为基底层角质形成细胞的损伤,不同程度的变性、液化坏死,以及固有层T淋巴细胞的带状浸润。2.口腔扁平苔藓患者病损区组织固有层及外周血中T淋巴细胞亚群的显著性改变,表明细胞免疫功能的紊乱在口腔扁平苔藓发病中发挥着重要作用。3.口腔黏膜上皮组织中角质形成细胞凋亡及凋亡抑制因子Bcl-2在病损区淋巴细胞浸润带处和凋亡促进因子Bax在上皮基底层角质形成细胞中表达增强,提示细胞凋亡异常与口腔扁平苔藓的发生、发展有重要关系,病损区组织超微结构观察结果也与之相符。4.TNF-α、IFN-γ在口腔扁平苔藓患者外周血中异常高表达,提示TNF-α、IFN-γ可能是造成口腔扁平苔藓病损的重要因子之一。
[Abstract]:(1) oral lichen planus (oral lichen planus, OLP) is the most common chronic noninfectious oral mucous membrane disease in clinic. It is a chronic inflammatory disease occurring in the skin and mucous membrane. By histopathological examination of 65 cases of oral lichen planus, the main pathological features are epithelial hyperkeratosis. The epithelium is irregular and accretion. The epithelium is irregular and serrated. The basal cell layer is liquefied and denatured, resulting in the disorder of the basal layer keratinocytes, the indistinct basement membrane boundaries, and the liquefaction and necrosis of the basal layer keratinocytes. The lamina is densely infiltrated with lymphocytes, the infiltrating zone is mainly distributed in the shallow layer of the lamina and part is also available. In 65 cases of oral lichen planus, 23 of the 65 cases of oral lichen planus were in the epithelia, and the basal layer or the lamina propria showed circular or oval shaped corpuscle. It was homogeneous, eosinophilic, and may be the product of apoptosis. The epithelia is mild atypical hyperplasia, showing epithelial cell differentiation. The cell size is different, the cell size is different, the morphology is different, the nucleus is large and dense, the proportion of the nuclear plasma is increased, the nuclear division can be increased but usually the normal nuclear division. The cell arrangement is disorderly, the cell level increases and the pole disappears. (2) CD4+, CD8+T cells in the oral lichen planus tissue and the CD8+T cells in 35 cases of oral lichen planus tissue and The expression level in the normal oral mucosa of 10 cases showed that the normal oral mucosa almost did not express CD4+, CD8+T cells, only the scattered CD4+, and positive expression of CD8+T cells in the basal layer. The oral lichen lamina propria showed an equal or large number of lymphocyte infiltration, and 92.3% (60/65) cases had lymphocytic infiltration. Oral squamous cell was flat. The expression level of CD8+ in the moss group was significantly higher than that in the control group (P0.01), and there was no significant difference between the CD4+ and the control group (P0.05), indicating that the occurrence of oral lichen planus may be related to the cellular immune changes in the pathological tissue. The CD8+T cells in the oral lichen planus group are mainly located in the superficial layer of the lamina propria, close to the expression of the basement membrane destruction, and the CD4+T cell master. There was a significant difference in the expression of.CD4/CD8 ratio in the deep lamina of the lamina propria (P0.0 5). (3) the expression level of Bcl-2 and Bax in the oral lichen planus and the control group was detected by immunohistochemical S-ABC. The results showed that Bcl-2 was mainly expressed in the cytoplasm and membrane of the upper layer and the acanthosis cells in the normal oral cavity. In the mucosa, 8/10 positive (+) expression, positive expression rate was 80%, in the epithelial tissue of oral lichen planus, 17/35 negative expression, 3/35 cases in the basal layer and spinous layer were scattered in the expression, 15/35 cases of lymphocyte infiltration zone positive expression increased, positive expression rate of 51.4%; Bcl-2 in oral lichen planus positive expression rate and normal oral mucus There was no significant change in the membrane (P0.05), but in the lymphocytic infiltration zone, there were 27 cases of.35 positive oral lichen planus. The positive rate was 27, the positive rate was 77.1%, of which 14 were positive (+ + +), 9 were moderately positive (+ +) and 4 were weak positive (+). The positive cells were the most obvious normal human oral mucus in the basal layer and the middle and lower part of the spine. In the membrane, 3 cases were weakly positive (+), and the other cases were negative (-). The positive expression rate of 30.0%.Bax in oral lichen planus was significantly higher than that in normal oral mucosa (P0.01). (4) the detection of pathological tissue in oral lichen planus and the cell withering in the control group by TUNEL (in situ terminal transferase labeling) The results showed that the apoptotic cells were expressed in the oral mucosa epithelium and lamina propria of 65 cases of oral lichen planus, with clear, dense, brown brown outline of the nucleus. The apoptotic index of the epithelial cells of the oral lichen planus was higher than that of the normal control group (P0.01), and the apoptotic cells were mostly located in the liquefied necrosis of the basal layer. The rate of arrival was 85.7% (30/35). The apoptotic cells in the control group were mostly located in the surface layer, spinous layer and basal layer, and the expression rate was 80% (8/10). (5) double anti sandwich enzyme-linked immunosorbent assay was used to detect the content of TNF-a and IFN- gamma in peripheral blood serum. The results showed that the content of IFN- gamma and TNF- alpha in the peripheral blood serum of the oral lichen planus patients was significantly higher than that of the control group (P0.05) (6). The T cell subgroup was identified and measured by the monoclonal antibody ring method. The results showed that compared with the healthy group, the CD8+ of 20 patients with oral lichen planus increased significantly (P0.05). There was no significant difference between the CD4+ and the control group (P0.05), and the CD4+/CD8+ decreased significantly (P0.01) compared with the control group (P0.01), and the results were consistent with the immunohistochemical staining analysis. (7) By transmission electron microscopy, it was found that the oral mucosa of oral lichen planus was filled with tension fibrils in the acanthosis of oral mucous membrane, and it became more and more coarse, arranged in disorder, the inner webs were dilated, there were unshaped objects, the mitochondria were enlarged and the crurtosis disappeared, the free ribosome increased, the majority of the organelles were reduced, the nucleus mass heterochromatin increased, and the internuclear gap was added. Wide, some paratype vacuoles, cell deformations, destruction of pbridge structure and obvious enlargement of intercellular space. Basal cells and basal membrane half bridge particles and intercellular pbridges, partial cell deformation, disorder, dislocation, enlargement of edema cell gap, vacuoles, denaturation, cytoplasmic swelling of endoplasmic reticulum, dilatation of endoplasmic reticulum, nucleus enlargement, heterostaining The number of apoptotic cells increased. The frequency of apoptotic cells was higher than that in the healthy group. Apoptosis was mainly manifested by the accumulation of chromatin along the border of the nuclear membrane, which was "C", crescent shape or mass, and apoptotic body. Conclusion the histopathological characteristics of 1. oral lichen planus are the damage of basal stratum corneum cells, varying degrees of degeneration and liquefaction. Death and zonal infiltration of the lamina propria T lymphocyte in.2. oral lichen planus patients the significant changes in the lamina propria and the T lymphocyte subsets in the peripheral blood, indicating that the dysfunction of the cellular immune function plays an important role in the pathogenesis of oral lichen planus and the apoptosis and apoptosis of keratinocytes in the oral mucosa of.3. oral mucosa The inhibitory factor Bcl-2 increased in the epithelial cell infiltration zone and the apoptotic factor Bax in the epithelial basal layer keratinocytes, suggesting that the abnormal apoptosis has an important relationship with the occurrence of oral lichen planus. The ultrastructural observation results of the lesion area also correspond to.4.TNF- a, IFN- gamma is in the oral lichen planus Abnormal high expression in peripheral blood indicates that TNF- alpha and IFN- gamma may be one of the important factors causing oral lichen planus lesions.
【学位授予单位】：郑州大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R781.5

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